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Related Experiment Video

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A Microfluidic Chip for the Versatile Chemical Analysis of Single Cells
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A Microfluidic Chip for the Versatile Chemical Analysis of Single Cells

Published on: October 15, 2013

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Microelectrophoretic single-cell measurements with microfluidic devices.

Jay Sibbitts1, Jalal Sadeghi1, Christopher T Culbertson1

  • 1Department of Chemistry, Kansas State University, Manhattan, KS, United States.

Methods in Enzymology
|November 1, 2019
PubMed
Summary
This summary is machine-generated.

We developed an automated microchip electrophoresis device for high-throughput single-cell analysis. This system accurately measures intracellular nitric oxide (NO) levels in T-lymphocytes, aiding research into chronic diseases.

Keywords:
ElectrophoreticEnzyme activityFiber opticLaser induced fluorescenceMicrochipMicrofluidicNitric oxideSingle cell analysis

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Area of Science:

  • Biomedical Engineering
  • Analytical Chemistry
  • Cell Biology

Background:

  • Single-cell analysis is crucial for understanding cellular heterogeneity and disease mechanisms.
  • Nitric oxide (NO) plays a vital role in various physiological processes and is implicated in chronic diseases.
  • Existing methods for intracellular NO measurement can be limited in throughput and precision.

Purpose of the Study:

  • To present the design, fabrication, and operation of an automated high-throughput single-cell microchip electrophoresis device.
  • To demonstrate the device's capability for precise intracellular nitric oxide (NO) level measurement in T-lymphocytes.
  • To enable adaptation of the system for diverse single-cell analyses.

Main Methods:

  • Development of a microchip electrophoresis device with integrated peristaltic pumps for precise fluid control.
  • Incorporation of an optical fiber bridge for simultaneous dual-point fluorescence detection.
  • Utilizing laser-induced fluorescence detection for sensitive analyte quantification.
  • Single-cell electropherogram generation with a t=0s reference signal for accurate migration time determination.

Main Results:

  • Successful fabrication and operation of the automated high-throughput single-cell microchip electrophoresis system.
  • Demonstrated accurate measurement of intracellular nitric oxide (NO) levels in T-lymphocytes.
  • Quantified NO levels under native, stimulated (lipopolysaccharide), and inhibited (inducible nitric oxide synthase) conditions.

Conclusions:

  • The developed microchip electrophoresis device offers a robust platform for high-throughput single-cell analysis.
  • The system provides a reliable method for quantifying intracellular NO, relevant to chronic disease research.
  • This technology has the potential to be adapted for a wide range of single-cell biomarker studies.